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Simplified Workflow For Hybridoma Antibody Sequencing
Before recombinant antibody expression plasmids can be designed, sequncing of the antibody light and heavy chain variable regions is necessary. Several other methods of sequencing antibody variable regions are available. Some involve high throughput RNA sequencing. These techniques are unavailable to many labs; they require the preparation of RNA-seq libraries, and computational analysis. As a result, the cost of performing such techniques is substantial and with sequencing cores being oversubscribed, turnaround can be as long as weeks to months. Other methods involve PCR and Sanger sequencing. However PCR amplification of variable regions results from difficulties in generating universal primers that can amplify any given variable region - particularly given the inherent low sequence identity in the 5' leader sequence of antibody light chains and heavy chains upstream of the variable regions. Sometimes degenerate primers can be used, but amplification success rate is only 80-90% due to non-specific priming and/or failure to prime at all. In addition, there is a significant risk that the variable regions of the parental myeloma line can amplify using the degenerate primers. 5' RACE (rapid amplification of 5' cDNA ends) can also be used, but mRNA degradation, cDNA purification and poly-A addition between reverse transcription and PCR, makes the technique long and difficult to perform. Non degenerate primers can be used, but each variable region requires multiple amplification attempts with different primer sets as well as sequence validation using mass spectrometry. And with both of these methods, primer derived mutations can be introduced. Mass spectrometry can be used to determine antibody variable regions, but these can result in ambiguous sequences because of isobaric residues such as isoleucine and leucine. But this method is time consuming, requires huge amounts of purified monoclonal antibody, is expensive and is inaccessible to most researchers. This technology involves a template switch reverse transcription of hybridoma RNA with at least three chain specific RT primers - one for the kappa chain, one for the lambda chain, and at least one for the heavy chain (for efficiency, this can be limited to IgG in a first pass). These are amplified in three separate PCR reactions and sequenced using Sanger sequencing.
Methods and Systems for Rapid Antimicrobial Susceptibility Tests
Rapid antimicrobial susceptibility testing (AST) is a method for quickly determining the most effective antibiotic therapy for patients with bacterial infections. These techniques enable the detection and quantification of antibiotic-resistant and susceptible bacteria metabolites at concentrations near or below ng/mL in complex media. Employing bacterial metabolites as a sensing platform, the system integrates machine learning data analysis processes to differentiate between antibiotic susceptibility and resistance in clinical infections within an hour. With the results, a clinician can prescribe appropriate medicine for the patient's bacterial infection.
CHD4 Targeting Peptide Isolated From Viral Protein For Cancer Therapeutics
Researchers at the University of California, Davis have identified a short peptide which rapidly promotes protein degradation in cancerous enzymes and induces cell differentiation to kill lymphomas.
Stable N-acetylated analogs of Sialic Acids and Sialosides
Researchers at the University of California, Davis have constructed a library of glycans containing N-acetyl sialic acids to mimic those containing naturally occurring O-acetyl sialic acids.
Anti-Human SULF2 monoclonal antibodies for research applications
Sulfatase 2 (SULF2) is an extracellular sulfatase that acts on heparan sulfate proteoglycans. It is overexpressed and pro-oncogenic in many cancers. Its overexpression in the liver is linked to dyslipidemia and fatty liver disease. This invention describes a panel of monoclonal antibodies that are validated for immunocytochemical staining, biochemical analysis and functional studies of human SULF2.
Monoclonal Antibodies Specific to Canine PD-1 and PD-L1
Researchers at the University of California, Davis have developed monoclonal antibodies with multiple applications relevant to canine PD-1 and PD-L1.
Kelch Like Family Member 11 (Klhl11) Autoantibodies As Markers Of Seminoma Associated Paraneoplastic Encephalitis In Men
Researchers at UCSF and Chan Zuckerberg Biohub have discovered a novel biomarker for an autoimmune disease that affects patients with testicular cancer. The disease, known as “testicular cancer-associated paraneoplastic encephalitis,” can cause severe neurological symptoms. The symptoms include loss of limb control, eye movement, and in some cases, speech. The disease begins with testicular cancer, which in some cases causes the immune system to attack the brain. Affected patients are often misdiagnosed and appropriate treatment is delayed.
New Indications For ENPP1 Inhibitors, Part Two
UCLA researchers in the Department of Medicine have developed small molecule ENPP1 inhibitors and monoclonal antibodies for treating myocardial infarction and ocular calcification.
Development of a New Biomarker for Diagnosis of Cardiovascular Disease: Monoclonal Antibody to Oxidized Cholesteryl Esters
Cardiovascular disease (CVD) is the leading cause of death and disability worldwide. The primary prevention of CVD is dependent upon the ability to identify high-risk individuals long before the development of overt events. This highlights the need for accurate risk stratification. An increasing number of novel biomarkers have been identified to predict cardiovascular events. Biomarkers play a critical role in the definition, prognostication, and decision-making regarding the management of cardiovascular events. There are several promising biomarkers that might provide diagnostic and prognostic information. The myocardial tissue-specific biomarker cardiac troponin, high-sensitivity assays for cardiac troponin, and heart-type fatty acid binding potential help diagnose myocardial infarction (MI) in the early hours following symptoms. Inflammatory markers such as growth differentiation factor-15, high-sensitivity C-reactive protein, fibrinogen, and uric acid predict MI and death and many others. However, there is a high unmet medical need for the more specific biomarkers that reflect different aspects of the development of atherosclerosis.
Disulfide Bioconjugation
UCLA researchers in the Department of Chemistry and Biochemistry have proposed a one-step radical mechanism for disulfide bioconjugation that overcomes many concerns associated with the free cysteine residues that result from current bioconjugation techniques.
Phenotypic Profiling Of Hepatocellular Carcinoma Circulating Tumor Cells For Treatment Selection
Researchers in the UCLA Departments of Surgery and Molecular and Medical Pharmacology have developed a novel blood-based assay that can capture and characterize circulating tumor cells indicative of both early- and late-staged hepatocellular carcinoma (HCC).
Anti-PSA IgG
Researchers at UCLA have developed chimeric IgG specific for prostate-specific antigen (PSA) in human and mouse cells.
Reagent to Label Proteins via Lysine Isopeptide Bonds
Researchers in the UCLA Department of Chemistry and Biochemistry and the University of Texas-Medical Center, Houston Department of Microbiology and Molecular Genetics have modified the Corynebacterium diphtheriae (C. diphtheriae) sortase enzyme so that it can be used as a bioconjugation reagent in vitro.
Targeting 3-Repeat TAU for the Treatment of Neurodegenerative Tauopathy Disorders
Neurodegenerative disorders with Tau accumulation are a common cause of dementia in the aging population. Alzheimer’s Disease (AD), Pick’s Disease (PiD) and Fronto-temporal lobar degeneration (FTLD) are examples of neurodegenerative disorders with Tau accumulation and are also jointly referred as “taupathies”. Tauopathies are a group of neurodegenerative disorders with accumulation of three-repeat (3R) or four-repeat (4R) Tau. While 3R tau is found in Pick's disease and Alzheimer's disease (AD), 4R tau is more abundant in corticobasal degeneration, progressive supranuclear palsy, and AD.
Antibodies for Pseudomonas (P.) aeruginosa
Researchers at the University of California, Davis have developed recombinant antibodies (VHH or nanobodies) to diagnose and treat Pseudomonas (P.) aeruginosa infection.
Sieve Container For Contactless Media Exchange For Cell Growth
Media that contains nutrients and growth factors is necessary to grow all types of cells, a process that is widely used in many fields of research. Such media should be routinely changed either to different media or a fresh batch of the same media. This change currently involves either using a pipette to transfer cells from their current dish of media to a new dish, or aspirating the media out of the dish and replacing it with new media. Both methods have inherent risks to stressing and damaging the cells. Researchers at UCI have developed a unique dish for growing cells that allows for safer aspiration of the old media, which reduces stress and damage to the cells.
Methods For Predicting Response Patterns To Anti-PD-1 (aPD-1) Therapy In Metastatic Melanoma
Dr. Roger Lo and colleagues in the Department of Medicine at UCLA have identified a method to predict a melanoma patient’s resistance to pembrolizumab and other immune checkpoint inhibitors.
Hybridoma Producing Antibodies To C1qRp
Individuals with genetic immunodeficiency, as well as patients with HIV, cancer, and those undergoing chemotherapy or high risk surgery, are at increased risk for infection. C1q, an important component of the immune system, is known to enhance phagocytosis (cell ingestion of harmful bacteria or other materials). Scientists at UCI have developed antibodies to the receptor for C1q, C1qRp, to be used as a target for prophylactic treatments in populations at high risk of infection.
Antibodies Against Human Mff Protein
UCLA researchers in the David Geffen School of Medicine Department of Biological Chemistry have developed antibodies against the human Mitochondrial fission factor (Mff).
Fusion Protein For Anti-Cd19 Chimeric Antigen Receptor Detection
Researchers at UCLA have developed a fusion protein that can detect immune cells expressing anti-CD19 chimeric antigen receptors with higher specificity and lower background than existing antibodies.
Treatment of spinal cord injury, traumatic brain injury, stroke and neurodegenerative disorders with a monoclonal antibody
Most people who suffer traumatic spinal cord injuries have incomplete lesions of neural circuits whose function can be partially restored from the reconfiguration of the spared circuits with rehabilitative training. Methods for improving nerve regeneration after spinal cord injury or nerve transplantation are needed for improved patient outcome. Also, neurodegenerative diseases such as amyotrophic lateral sclerosis, Alzheimer’s Disease and Parkinson’s Disease negatively impact quality of life.
Pyrite Shrink-Wrap Laminate As A Hydroxyl Radical Generator
The invention is a diagnostic technology, as well as a research and development tool. It is a simple, easy to operate, and effective platform for the analysis of pharmaceuticals and biological species. Specifically, this platform generates hydroxyl radicals for oxidative footprinting – a technique commonly employed in protein mapping and analysis. The platform itself is inexpenisve to fabricate, scalable, and requires nothing more than an ordinary pipet to use. In addition, it is highly amenable to scale-up, multiplexing, and automation, and so it holds promise as a high-throughput method for mapping protein structure in support of product development, validation, and regulatory approval in the protein-based therapeutics industry.
Monoclonal Antibody Against PNPase (Clone 4C11)
Mouse monoclonal antibody against the human mitochondrial polyribonucleotide nucleotidyltransferase 1 (PNPase). This antibody has been tested for use in immunocytochemistry/immunofluorescence, immunoprecipitation, and western blot.
Antibodies for the Detection of Toxoplasma Gondii Oocysts
Researchers at the University of California, Davis have developed monoclonal antibodies that recognize and bind to oocysts of Toxoplasma gondii.
Immunomodulary Materials for Implantable Medical Devices
The host response to biomaterials is a huge challenge for the design of medical devices. Researchers at UC Irvine have created an extremely biocompatible material that can escape inflammatory immune attack.